雷群 陈江 林恒章 黄文秀 吴东 杨静.磷离子对 BM-hMSCs 增殖及成骨分化的影响[J].现代生物医学进展英文版,2014,14(35):6839-6842. |
磷离子对 BM-hMSCs 增殖及成骨分化的影响 |
Study on the Effect of P Ion on the Proliferation and OsteogenicDifferentiation of BM-hMSCs |
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DOI: |
中文关键词: 磷离 子 骨髓间充质干细胞 增殖 成骨分化 |
英文关键词: P Ion Bone marrow mesenchymal stem cells Proliferation Osteogenic differentiation |
基金项目:2012 年福建省临床重点专科项目 资助(闽卫科教[2012]1 49 号) |
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中文摘要: |
目的: 探讨磷离 子对人骨髓来源的间充质干细胞(Human bone marrow mesenchymal stem cells, BM-hMSCs)增殖及成骨分化
的影响。 方法: 从 ScienCell 实验室购买的 BM-hMSCs 分别在无血清生长培养基(对照组)和添加 4mmol(4P 组)、 8mmol(8P 组)磷离
子的无血清生长培养基中培养 21 天, 通过 CCK8 比色法评估细胞的增殖情况; RT-PCR 检测成骨分化标记性基因 胶原蛋白 Ⅰ 、骨
钙素、碱性磷酸酶的表达水平; 茜素红染色检测 BM-hMSCs 成骨分化产生的矿化结节。 结果: 在培养 4、 7、 14 天时, 4P 和 8P 组中
BM-hMSCs 的增殖都明显高于对照组, 且 8P 组中 BM-hMSCs 的增殖高于 4P 组; 培养 21 天时, 4P 和 8P 组中 BM-hMSCs 的增殖
明显低于对照组。 与 对照组相比,在培养 7 天时, 4P 和 8P 组 OC 的表达下调,而 14、 21 天时, OC 的表达上调。 4P 和 8P 组中 ALP
的表达水平与 对照组无明显差异。 7 天时, 4P、 8P 组 ColⅠ 的表达水平均明显高与 对照组; 14 天时, 8P 组 ColⅠ 的表达水平与 对照
组表达无明 显差异; 21 天时, 4P、 8P 组中 ColⅠ 的表达水平都均与 对照组表达无明 显差异。 培养 21 天时, 4P、8P 组矿化结节的形
成明显增加。 结论: 磷离 子在早期可以促进 BM-hMSCs 的增殖, 晚期诱导其成骨分化。 |
英文摘要: |
Objective:To investigate the effect of P ion on proliferation and differentiation of BM-hMSCs.Methods:BM-hMSCs
purchased from ScienCell Research Laboratories were subjected to serum free growth medium (control group) and serum free growth
medium with additional 4mmol Pi(4P) or 8mmol Pi(8P) for 21 d. CCK8 colorimetric method was used to assess the cellular proliferation;
RT-PCR was used to examine the mRNA expression of osteogenic marker genes, including alkaline phosphatase, collagen typeⅠ and
osteocalcin; Alizarin Red-S method was used to study the mineralization of BM-hMSCs.Results:The proliferation of BM-hMSCs
significantly increased in 4P and 8P groups in day 4, 7 and 1 4 whereas decreased in day 21 compared to that in the control group. The
proliferation of BM-hMSCs was more obvious in 8P group. The mRNA expression of OC was down-regulated in 4P and 8P groups in
day 7 whereas the mRNA expression of OC was up-regulated in day 1 4 and 21 compared to control group.The mRNA expression of ALP
in 4P and 8P group was not different with that in control group. The mRNA expression of collagen typeⅠ significantly increased in 4P
group in day 7 and 14, but only increased in 8P group in day 7. The formation of mineralization nodules in BM-hMSCs were obvious in
both 4P and 8P groups in day 21.Conclusion:Our data indicated that Pi could promote proliferation at an early stage and induced
osteogenic differentiation of BM-hMSCs.at late stage. |
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